Multi-component formulations

ABSTRACT

Multi-component formulations comprise methylsulfonylmethane, fructose 1,6-diphosphate, and just one of trehalose, green tea extract, ashwagandha, and rutin, or a combination thereof. Multi-component formulations can prevent, delay, or treat cognitive decline, Alzheimer&#39;s disease, and other dementias.

FIELD OF INVENTION

This invention relates to compositions for administration to human or animal patients, and methods of preventing, delaying, and treating cognitive decline, Alzheimer's disease and other dementias, and other neurodegenerative diseases using those compositions.

BACKGROUND OF THE INVENTION

Alzheimer's disease (“AD”) is believed to be a multifactorial degenerative disease rather than the result of a single malfunction or agent. Although AD is usually accompanied by the abnormal accumulation of extracellular deposits or plaques of β-amyloid protein (AB) and intracellular neurofibrillary tangles of tau protein (NFTs), extensive research has not isolated or identified a cause for the accumulation, nor shown that β-amyloid protein or NFTs are the cause rather than effect of AD. In fact, approximately 30 percent of AD patients have no AB plaques or NFTs at death, and approximately 30 percent of cognitively normal adults do have AB plaques and NFTs at death. AB plaques may even be protective against harmful soluble AB oligomers. Instead, clinical and epidemiological studies have identified numerous contributing factors to AD.

Many of the pathologic characteristics of AD, inflammation, oxidative stress, impaired cerebral blood flow and glucose utilization, result from body imbalances, including stress, obesity, and an overloaded immune system. In addition, AD has a lengthy, non-linear, accelerating, and degenerative prodromal time period with ample opportunity for preventive intervention, and about 95 percent of AD cases are sporadic late-onset.

However, no comprehensively-effective medicine or protocol has emerged for AD or other dementias. Yet it is estimated that about 44 million patients suffer from AD or a related dementia worldwide. Cognitive decline affects significantly more. New compositions designed on the growing understanding of neurobiochemistry, cognitive decline, AD, other dementias, and neurodegeneration in general are needed.

SUMMARY OF THE INVENTION

Unexpectedly, Applicant has invented certain compositions that can be used in the prevention, delay, and treatment of cognitive decline, such as, for example, in AD patients. Accordingly, some embodiments of the present invention relate to multi-component formulations comprising methylsulfonylmethane, fructose 1,6-diphosphate, and at least one of trehalose, green tea extract, ashwagandha, and rutin. In certain instances, the multi-component formulations comprise just one of trehalose, green tea extract, ashwagandha, and rutin.

Further embodiments relate to methods of preventing or delaying cognitive decline in a human or animal patient in need thereof, one such method comprising administering to the patient an effective amount of a multi-component formulation comprising methylsulfonylmethane, fructose 1,6-diphosphate, and at least one of trehalose, green tea extract, ashwagandha, and rutin. In certain instances, the multi-component formulation comprises just one of trehalose, green tea extract, ashwagandha, and rutin.

Additional embodiments relate to methods of preventing or delaying AD or other dementia in a human or animal patient in need thereof, one such method comprising administering to the patient an effective amount of a multi-component formulation comprising methylsulfonylmethane, fructose 1,6-diphosphate, and at least one of trehalose, green tea extract, ashwagandha, and rutin. In certain instances, the multi-component formulations comprise just one of trehalose, green tea extract, ashwagandha, and rutin.

Yet other embodiments relate to methods of treating cognitive decline in a human or animal patient in need thereof, comprising administering to the patient an effective amount of a multi-component formulation comprising methylsulfonylmethane, fructose 1,6-diphosphate, and at least one of trehalose, green tea extract, ashwagandha, and rutin. In certain instances, the multi-component formulation comprises just one of trehalose, green tea extract, ashwagandha, and rutin.

Still additional embodiments relate to methods of treating AD or other dementia in a human or animal patient in need thereof, comprising: administering to the patient an effective amount of the multi-component formulation comprising methylsulfonylmethane, fructose 1,6-diphosphate, and at least one of trehalose, green tea extract, ashwagandha, and rutin. In certain instances, the multi-component formulation comprises just one of trehalose, green tea extract, ashwagandha, and rutin.

Yet further additional embodiments provide methods of making the multi-component formulation of any one of the preceding claims, one such method comprising combining methylsulfonylmethane, fructose 1,6-diphosphate, and at least one of trehalose, green tea extract, ashwagandha, and rutin into a form suitable for administration to a human or animal patient in need thereof. In certain instances, the multi-component formulation comprises just one of trehalose, green tea extract, ashwagandha, and rutin.

While the disclosure provides certain specific embodiments, the invention is not limited to those embodiments. A person of ordinary skill will appreciate from the description herein that modifications can be made to the described embodiments and therefore that the specification is broader in scope than the described embodiments. All examples are therefore non-limiting.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts the effect on cell viability of human neural stem cell derived neurons of a composition comprising methylsulfonylmethane, fructose 1,6-diphosphate, and ashwagandha.

FIG. 2 depicts the effect on cell viability of human neural stem cell derived neurons of a composition comprising methylsulfonylmethane, fructose 1,6-diphosphate, and green tea extract.

FIG. 3 depicts the effect on cell viability of human neural stem cell derived neurons of a composition comprising methylsulfonylmethane, fructose 1,6-diphosphate, and trehalose.

FIG. 4 depicts the effect on cell viability of human neural stem cell derived neurons of a composition comprising chlorpromazine.

FIG. 5 depicts the effect on cell viability of primary chicken neurons of a composition comprising methylsulfonylmethane.

FIG. 6 depicts the effect on cell viability of primary chicken neurons of a composition comprising fructose 1,6-diphosphate.

FIG. 7 depicts the effect on cell viability of primary chicken neurons of a composition comprising 1 mM methylsulfonylmethane and varying concentrations of fructose 1,6-diphosphate.

FIG. 8 depicts the effect on cell viability of primary chicken neurons of a composition comprising 0.1 mM methylsulfonylmethane and varying concentrations of fructose 1,6-diphosphate.

FIG. 9 depicts the effect on cell viability of primary chicken neurons of a composition comprising 0.01 mM methylsulfonylmethane and varying concentrations of fructose 1,6-diphosphate.

FIG. 10 depicts the effect on cell viability of primary chicken neurons of a composition comprising 0.001 mM methylsulfonylmethane and varying concentrations of fructose 1,6-diphosphate.

FIG. 11 depicts the effect on cell viability of primary chicken neurons of a composition comprising 0.0001 mM methylsulfonylmethane and varying concentrations of fructose 1,6-diphosphate.

FIG. 12 depicts the effect on cell viability of primary chicken neurons of compositions comprising a vehicle control (“VC”), methylsulfonylmethane and fructose 1,6-diphosphate (“MF”), and methylsulfonylmethane, fructose 1,6-diphosphate, and rutin (“MFR”).

FIG. 13 depicts the effect on cell viability of human neural stem cell derived neurons of various compositions including those containing various concentrations of rutin trihydrate.

DETAILED DESCRIPTION

As required, detailed embodiments of the present invention are disclosed herein; however, it is to be understood that the disclosed embodiments are merely exemplary of the invention that may be embodied in various forms. The figures are not necessarily to scale, and some features may be exaggerated to show details of particular components. Therefore, specific structural and functional details disclosed herein are not to be interpreted as limiting, but merely as a basis for the claims and as a representative basis for teaching one skilled in the art to variously employ the present invention.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of ordinary skill in the art to which this disclosure belongs. In the event that there is a plurality of definitions for a term herein, those in this section prevail unless stated otherwise.

Where ever the phrase “for example,” “such as,” “including” and the like are used herein, the phrase “and without limitation” is understood to follow unless explicitly stated otherwise. Similarly “an example,” “exemplary” and the like are understood to be non-limiting.

The term “substantially” allows for deviations from the descriptor that don't negatively impact the intended purpose. Descriptive terms are understood to be modified by the term “substantially” even if the word “substantially” is not explicitly recited.

The term “about” when used in connection with a numerical value refers to the actual given value, and to the approximation to such given value that would reasonably be inferred by one of ordinary skill in the art, including approximations due to the experimental and or measurement conditions for such given value.

The terms “comprising” and “including” and “having” and “involving” (and similarly “comprises”, “includes,” “has,” and “involves”) and the like are used interchangeably and have the same meaning. Specifically, each of the terms is defined consistent with the common United States patent law definition of “comprising” and is therefore interpreted to be an open term meaning “at least the following,” and is also interpreted not to exclude additional features, limitations, aspects, etc. Thus, for example, “a device having components a, b, and c” means that the device includes at least components a, b and c. Similarly, the phrase: “a method involving steps a, b, and c” means that the method includes at least steps a, b, and c.

Unless the context clearly requires otherwise, throughout the description and the claims, the words “comprise”, “comprising”, and the like are to be construed in an inclusive sense as opposed to an exclusive or exhaustive sense; that is to say, in the sense of “including, but not limited to”.

Any discussion of the prior art throughout the specification should in no way be considered as an admission that such prior art is widely known or forms part of common general knowledge in the field.

It is an object of the present invention to overcome or ameliorate at least one of the disadvantages of the prior art, or to provide a useful alternative.

In view of the numerous and substantial benefits to be realized from formulations exhibiting efficacy in the prevention, delay, and treatment of one or more of the wide variety of degenerative effects which accompany AD, the present invention is directed to unique new multi-component formulations for the prevention, delay, and/or treatment of AD, cognitive decline, and/or other neurodegenerative diseases, which counters the multiple contributing factors to AD, cognitive decline, and/or other neurodegenerative diseases, with a novel combination of complementary remedies.

The complementary components combine drugs and herbals, Eastern and Western thinking, natural molecules and nutrients. All of the components of the present formulation, individually and together, work with the body and brain in order to help the body and brain themselves prevent AD. The present formulation should be combined with proper diet, sleep, exercise without stress, also the best regulator of insulin sensitivity, and social/mental/spiritual outlook and lifestyle, including building up “cognitive reserve,” in order to minimize the uniquely human and degenerative nature of AD. Although the human brain makes up only 2 percent of the body's mass, it is an organ of amazing complexity and design which controls the entire body. It has virtually no stored oxygen or glucose, and consumes 20 percent of the body's oxygen and 25 percent of the body's glucose, its sole source of energy. As a result, the human brain is continually on the brink of hypoxia and has an enormous and constant appetite for blood and energy, specifically, as much as 15 percent of the blood and 25 percent of the body's essential energy supply at any moment. Oxygen-glucose deprivation then leads to mitochondrial dysfunction and stimulation of neuroprotective proteins. Many of the pathologic characteristics of AD—inflammation, oxidative stress, impaired cerebral blood flow and glucose utilization—result from body imbalances, including stress, obesity, and an overloaded immune system.

Certain formulations of the present invention can help to restore the body's proper balance, and proper lifestyle can help to maintain that balance. The present invention provides a multi-component formulation for the prevention, delay, and/or treatment of AD, cognitive decline, and/or other neurodegenerative diseases.

As stated above, certain embodiments of the present invention relate to a multi-component formulation comprising: methylsulfonylmethane, fructose 1,6-diphosphate, and just one or more than one of trehalose, green tea extract, ashwagandha, and rutin. Any suitable composition can be used as the formulations of the present invention. The term formulation shall include liquids, semi-liquids, colloidal solutions, dispersions, emulsions, microemulsions, and nanoemulsions, including oil-in-water emulsions and water-in-oil emulsions, pastes, powders, and suspensions. The formulations of the present invention may also be included, or packaged, with other non-toxic compounds, such as cosmetic carriers, excipients, binders and fillers, and the like. Specifically, the acceptable cosmetic carriers, excipients, binders, and fillers contemplated for use in the practice of the present invention are those which render the compounds amenable to oral delivery and/or provide stability such that the formulations of the present invention exhibit a commercially acceptable storage shelf life.

Certain instances of the present invention relate to methods of making the multi-component formulations described herein, one such method comprising combining the methylsulfonylmethane, fructose 1,6-diphosphate, and the at least one of trehalose, green tea extract, ashwagandha, and rutin into a form suitable for administration to a human or animal patient in need thereof. The compositions of the invention can be brought into forms suitable for administration by means of usual processes optionally using auxiliary substances such as liquid or solid, powdered ingredients, such as the pharmaceutically customary liquid or solid fillers and extenders, solvents, emulsifiers, lubricants, flavorings, colorings and/or buffer substances. Frequently used auxiliary substances include magnesium carbonate, titanium dioxide, lactose, saccharose, sorbitol, mannitol and other sugars or sugar alcohols, talc, lactoprotein, gelatin, starch, amylopectin, cellulose and its derivatives, animal and vegetable oils such as fish liver oil, sunflower, groundnut or sesame oil, polyethylene glycol and solvents such as, for example, sterile water and mono- or polyhydric alcohols such as glycerol, as well as with disintegrating agents and lubricating agents such as magnesium stearate, calcium stearate, sodium stearyl fumarate and polyethylene glycol waxes. The mixture may then be processed into granules or pressed into tablets.

The active ingredients may be separately premixed with the other non-active ingredients, before being mixed to form a formulation. The active ingredients may also be mixed with each other, before being mixed with the non-active ingredients to form a formulation.

Soft gelatin capsules may be prepared with capsules containing a mixture of the active ingredients of the invention, and optionally vegetable oil, fat, or other suitable vehicle for soft gelatin capsules. Hard gelatin capsules may contain granules of the active ingredients. Hard gelatin capsules may also contain the active ingredients together with solid powdered ingredients such as lactose, saccharose, sorbitol, mannitol, potato starch, corn starch, amylopectin, cellulose derivatives or gelatin.

Liquid preparations may be prepared in the form of syrups, elixirs, concentrated drops or suspensions, e.g. solutions or suspensions containing the active ingredients and the remainder consisting, for example, of sugar or sugar alcohols and a mixture of ethanol, water, glycerol, propylene glycol and polyethylene glycol. If desired, such liquid preparations may contain coloring agents, flavoring agents, preservatives, saccharine and carboxymethyl cellulose or other thickening agents. Liquid preparations may also be prepared in the form of a dry powder, reconstituted with a suitable solvent prior to use. Solutions for parenteral administration may be prepared as a solution of a formulation of the invention in a pharmaceutically acceptable solvent. These solutions may also contain stabilizing ingredients, preservatives and/or buffering ingredients. Solutions for parenteral administration may also be prepared as a dry preparation, reconstituted with a suitable solvent before use.

Certain embodiments comprise methylsulfonylmethane, fructose 1,6-diphosphate, and trehalose, and no other active ingredient. Other embodiments comprise methylsulfonylmethane, fructose 1,6-diphosphate, and green tea extract, and no other active ingredient. Yet additional embodiments comprise methylsulfonylmethane, fructose 1,6-diphosphate, and ashwagandha, and no other active ingredient. Still other embodiments comprise methylsulfonylmethane, fructose 1,6-diphosphate, and rutin, and no other active ingredient.

Methylsulfonylmethane, fructose 1,6-diphosphate, trehalose, green tea extract, ashwagandha, and rutin have the broadest suitable meanings as understood by one of ordinary skill in the art. Methylsulfonylmethane, (CH₃)₂SO₂, is also known as dimethyl sulfone. Fructose 1,6-diphosphate is also known as fructose 1,6-bisphosphate. Any suitable cations can be used to satisfy the negative charges centered on the phosphate groups, such as, for example, protons, alkali metal cations, alkaline earth metal cations, transition metal cations, nitrogen-containing cations, and combinations thereof. The naturally-occurring enantiomer appears in some embodiments of the present invention. D-fructose 1,6-diphosphate trisodium salt hydrate can be mentioned for some embodiments. Trehalose is the disaccharide also known as mycose and tremalose. D-(+)-trehalose anhydrous can be mentioned in certain instances. Green tea extract indicates any substance extracted from the leaves or any portion of the Camellia sinensis plant by any suitable methods. Fresh or dried leaves can be ground, immersed in any suitable solvent such as water, ethanol, or the like, brought to reflux, and then the solvent can be evaporated, leaving a green tea extract, for example. The naturally-occurring enantiomer appears in certain embodiments of the present invention. Ashwagandha refers to any portion or extract from the plant Withania somnifera. Roots and extracts thereof are mentioned. Rutin is also known as quercetin-3-O-rutinoside. The naturally-occurring enantiomer appears in further embodiments of the present invention. Rutin trihydrate can be mentioned for certain embodiments.

The active ingredients can be present in the multi-component formulations in any suitable amount. In some cases, for example, methylsulfonylmethane comprises at least 0.01%, 0.01% to 0.5%, 0.5% to 1.0%, 1.0% to 5.0%, 5.0% to 10%, 10% to 20%, 20% to 50%, 50% to 75%, 75% to 95%, or less than 99% by weight of the formulation. In other cases, fructose 1,6-diphosphate comprises at least 0.01%, 0.01% to 0.5%, 0.5% to 1.0%, 1.0% to 5.0%, 5.0% to 10%, 10% to 20%, 20% to 50%, 50% to 75%, 75% to 95%, or less than 99% by weight of the formulation. Additional cases provide trehalose in at least 0.01%, 0.01% to 0.5%, 0.5% to 1.0%, 1.0% to 5.0%, 5.0% to 10%, 10% to 20%, 20% to 50%, 50% to 75%, 75% to 95%, or less than 99% by weight of the formulation. In yet further cases, green tea extract is present as at least 0.01%, 0.01% to 0.5%, 0.5% to 1.0%, 1.0% to 5.0%, 5.0% to 10%, 10% to 20%, 20% to 50%, 50% to 75%, 75% to 95%, or less than 99% by weight of the formulation. Yet other cases provide ashwagandha as at least 0.01%, 0.01% to 0.5%, 0.5% to 1.0%, 1.0% to 5.0%, 5.0% to 10%, 10% to 20%, 20% to 50%, 50% to 75%, 75% to 95%, or less than 99% by weight of the formulation. Further additional cases provide rutin in an amount of at least 0.01%, 0.01% to 0.5%, 0.5% to 1.0%, 1.0% to 5.0%, 5.0% to 10%, 10% to 20%, 20% to 50%, 50% to 75%, 75% to 95%, or less than 99% by weight of the formulation.

Some embodiments of the present invention relate to preventing, delaying, or treating cognitive decline or AD in a human or animal patient in need thereof. Those methods comprise administering to the patient an effective amount of the multi-component formulations described herein. Any suitable effective amount can be employed. As used herein, “effective amount” means the amount necessary to render the desired therapeutic result. For example, an effective amount is a level effective to treat, cure, or alleviate the symptoms of a disorder for which the therapeutic compound, biologic or composition is being administered. Amounts effective for the particular therapeutic goal sought will depend upon a variety of factors including the disorder being treated and its severity and/or stage of development/progression; the bioavailability, and activity of the specific compound or formulation used; the route or method of administration and introduction site on the subject; the rate of clearance of the specific compound or biologic and other pharmacokinetic properties; the duration of treatment; inoculation regimen; drugs used in combination or coincident with the specific composition or formulation; the age, body weight, sex, diet, physiology and general health of the subject being treated; and like factors well known to one of skill in the relevant scientific art. Some variation in dosage will necessarily occur depending upon the condition of the patient being treated, and the physician or other individual administering treatment will, in any event, determine the appropriate dose for an individual patient. Thus, it is not useful to specify an exact effective amount in advance. However, in some cases, a dose of a multi-component formulation according to the present invention can be on the order of less than 10 μg, about 10 μg, about 100 μg, about 1 mg, about 10 mg, about 100 mg, about 250 mg, about 500 mg, about 1 g, or more than 1 g.

As used herein, the term “administering” refers to providing a therapeutically effective amount of a formulation or pharmaceutical composition to a subject, using intravitreal, intraocular, ocular, subretinal, intrathecal, intravenous, subcutaneous, transcutaneous, intracutaneous, intracranial, topical and the like administration. The formulation or pharmaceutical compound of the present invention can be administered alone, but may be administered with other compounds, excipients, fillers, binders, carriers or other vehicles selected based upon the chosen route of administration and standard pharmaceutical practice. Administration may be by way of carriers or vehicles, such as injectable solutions, including sterile aqueous or non-aqueous solutions, or saline solutions; creams; lotions; capsules; tablets; granules; pellets; powders; suspensions, emulsions, or microemulsions; patches; micelles; liposomes; vesicles; implants, including microimplants; eye drops; other proteins and peptides; synthetic polymers; microspheres; nanoparticles; and the like.

The formulations or pharmaceutical composition of the present invention may also be included, or packaged, with other non-toxic compounds, such as pharmaceutically acceptable carriers, excipients, binders and fillers including, but not limited to, glucose, lactose, gum acacia, gelatin, mannitol, xanthan gum, locust bean gum, galactose, oligosaccharides and/or polysaccharides, starch paste, magnesium trisilicate, talc, corn starch, starch fragments, keratin, colloidal silica, potato starch, urea, dextrans, dextrins, and the like. Specifically, the pharmaceutically acceptable carriers, excipients, binders, and fillers contemplated for use in the practice of the present invention are those which render the compounds of the invention amenable to intravitreal delivery, intraocular delivery, ocular delivery, subretinal delivery, intrathecal delivery, intravenous delivery, subcutaneous delivery, transcutaneous delivery, intracutaneous delivery, intracranial delivery, topical delivery and the like. Moreover, the packaging material may be biologically inert or lack bioactivity, such as plastic polymers, silicone, etc. And may be processed internally by the subject without affecting the effectiveness of the composition/formulation packaged and/or delivered therewith.

As used herein, “treatment” or “treating” refers to arresting or inhibiting, or attempting to arrest or inhibit, the progression of a disorder and/or causing, or attempting to cause, the reduction, suppression, regression, or remission of a disorder and/or a symptom thereof. As would be understood by those skilled in the art, various clinical and scientific methodologies and assays may be used to assess the development or progression of a disorder, and similarly, various clinical and scientific methodologies and assays may be used to assess the reduction, regression, or remission of a disorder or its symptoms.

As used herein, “prevention” or “preventing” refers to arresting or inhibiting, or attempting to arrest or inhibit, the initial onset or development of a disorder. The prevention of a disorder may occur before any clinical signs of the disorder being prevented are recognized.

In one embodiment, the disorder being treated, delayed, and/or prevented by the formulations and compositions of the present invention is a neurodegenerative disease, such as dementia or cognitive decline, which includes at least AD.

The many contributing factors to AD, together with each individual's unique makeup, genetic and otherwise, create a mind-boggling number of combinations which the present multi-component formulation is uniquely qualified to counter. If one component does not work with a particular individual, another component may. The multiple synergies between the different components of the present formulation increase the opportunity for at least one such synergy to provide an effective treatment for a particular person, unlike conventional single component drugs, which fail to benefit one in three patients and make one in seven to eight worse.

Different forms of the present inventive formulation can be calibrated in order to adapt both to different individuals and to the different needs of a single individual. Implementing this concept is complicated, and the necessary research is challenging. However, the present formulation need not counter every cause in every individual. Rather, by countering the necessary causes, the present formulation will restore the body and brain to their normal function. Then the body and brain themselves will correct the remaining deficiencies. No drug can possibly correct every single cause of AD, but the present formulation will maximize the possibility.

The end result of the present inventive formulation is to push back the threshold age of development of AD or cognitive decline, i.e., to delay or prevent the onset of AD until later in life, or alternatively, to eliminate AD or cognitive decline altogether for some people. The formulation is not expected to be a cure for advanced AD. AD has a lengthy, non-linear, accelerating, and degenerative prodromal time period with ample opportunity for preventive intervention. Approximately 95% of AD cases are sporadic late-onset. The inventive formulation should be combined with early detection of mild cognitive impairment (MCI) and cognitive impairment which is not dementia (CIND), conditions that may develop into AD, in order to maximize its effectiveness. MCI/CIND is a fluid, non-homogeneous, and unstable state, which is a critical time to intervene. The inventive formulation may be marketed to all people over 50 as a preventive measure, including those with no cognitive impairment. A 5-year delay of the onset of AD would yield spectacular results.

Testing of formulations of the present invention can be performed in accordance with any suitable protocol. Some such testing could be focused on maximizing the exogenous stimulation of endogenous neuroprotective proteins, mild stress-induced hormesis, inhibiting the production of reactive oxygen species, restoring homeostasis, and on examining the interrelationships and synergies between such neuroprotective proteins, the immune system, DNA (including SIRT1), epigenetics, cell senescence, neurogenesis, and neurodegeneration.

Without further elaboration, it is believed that one skilled in the art can utilize the present invention to its fullest extent based on the disclosure herein. The following examples are offered by way of illustration, not by way of limitation. While specific examples have been provided, the above description is illustrative and not restrictive. Any one or more of the features of the previously described embodiments can be combined in any manner with one or more features of any other embodiments in the present invention. Furthermore, many variations of the invention will become apparent to those skilled in the art upon review of the specification.

EXAMPLES Example 1—Cell Viability of Human Stem Cell Derived Neurons Treated with Multi-Component Formulations

Cryopreserved human stem cell derived neurons obtained from PhoenixSongs Biologicals were thawed and plated at a density of 12000 cells per well in a 384-well PDL/laminin coated plate. The neurons were maintained in a humidified environment at 37° C. with 5% CO₂ for six days with periodic media changes before experimental procedures were performed. The media used for maintenance and dosing was PhoenixSongs Neural Differentiation Medium and for the survival test contained methylsulfonylmethane (“MSM”) and fructose 1,6-diphosphate (“FDP”) (specifically, D-fructose 1,6-diphosphate trisodium salt hydrate) with final concentrations of 0.1 μM and 1 μM, respectively. Cells were dosed in triplicate with 10-point dose response curves to determine if there was any protective effect (improved survival of cells) or toxicity with the test compounds versus the untreated cells. A positive control (chlorpromazine) was also used to assess cytotoxicity response in these cells.

After a three-day incubation with test compositions, the cells were stained with Hoechst and propidium iodide and read on an ArrayScan VTI to determine cell viability. Results were normalized to vehicle and graphed using GraphPad Prism. FIG. 1 shows the dose response curve for ashwagandha; FIG. 2 shows the dose response curve for green tea extract known as Teavigo (90% purity); FIG. 3 shows the dose response curve for D-(+)-trehalose anhydrous; and FIG. 4 shows the dose response curve for chlorpromazine (without methylsulfonylmethane and fructose 1,6-diphosphate). IC50 values appear in Table 1 below:

TABLE 1 Formulation IC₅₀ for Cell Loss MSM + FDP + Ashwagandha >50 μg/mL MSM + FDP + Green Tea Extract >20 μM MSM + FDP + Trehalose >1000 μM Chlorpromazine 3.8 μM

Chlorpromazine is used as an antipsychotic in spite of its relatively high neuron cell toxicity as shown in this study. The inventive compositions are shown to be much safer at comparable concentrations than the standard antipsychotic compound.

Example 2—Cell Viability of Primary Chicken Neurons Treated with Rutin

Fertilized chicken eggs were stored under turning at 37.8° C. and 55% humidity until embryonic day eight. Embryos were transferred to a plastic dish and decapitated. Both hemispheres were removed, collected and cleaned from any loose tissue. Hemispheres were mechanically dissociated and 4.8×10⁴ cells per well (96-well plates) were seeded in a volume of 160 μL. The cell culture medium consisted of Dubelcco's Modified Eagle's Medium with 4.5 g glucose, 5% Nu Serum, 0.01% gentamycin and 2 mM L-glutamine. Cultures were maintained at 37° C., 95% humidity and 5% CO₂.

Test compositions and vehicle controls were applied to chicken neurons at day-in-vitro 8 for 48 h. On day-in-vitro 10 the viability of neurons was determined according to MTT cell viability assay using a plate reader (570 nm). That assay allows the measurement of the mitochondrial dehydrogenase activity, which reduces yellow MTT to dark blue formazan crystals. Since this reaction is catalyzed in living cells only, this assay is used for the determination of cell viability. MTT solution was added to each well in a final concentration of 0.5 mg/mL. After two hours the MTT-containing medium is aspired. Cells were lysed in 3% SDS and the formazan crystals were dissolved in isopropanol/HCl. Optical density was measured with a plate reader at 570 nm. Cell survival rage was expressed as optical density (OS). Values were calculated as percent of control values (vehicle control, which is medium).

Results from administering rutin in vehicle alone appear in Table 2:

TABLE 2 Rutin VC 0.001 0.01 0.1 1 10 Mean 100.0 96.7 99.7 98.1 97.7 100.2 Std. Deviation 3.97 6.08 8.57 4.79 7.91 9.21 Std. Error 1.15 1.76 2.47 1.38 2.39 2.66

Application of rutin (with no MSM nor FDP) did not enhance cell viability compared to vehicle-treated cells.

Results from administering MSM, FDP, and rutin appear in Table 3:

TABLE 3 Rutin + MSM/FDP VC MF 0.001 0.01 0.1 1 10 Mean 100.0 97.1 108.0 103.4 104.2 100.2 104.1 Std. Deviation 6.37 0.01 7.04 6.48 9.60 9.89 4.45 Std. Error 1.84 3.04 2.03 1.87 2.77 2.85 1.29

A combination of methylsulfonylmethane, fructose 1,6-diphosphate, and rutin at a concentration of 0.001 μM significantly improved cell viability compared to vehicle or MSM/FDP-only treated cells. At other concentrations, MSM/FDP/Rutin improved cell viability compared to MSM/FDP alone in this study.

Example 3—MSM and FDP Alone or Together

Using the same protocol as in Example 2, primary chicken neurons were treated with varying concentrations of methylsulfonylmethane and/or fructose 1,6-diphosphate. Cell viability was determined with the MTT assay. Compared to vehicle control, FIG. 5 shows that cell viability declined at MSM concentrations above 5 mM. FIG. 6 shows that cell viability declined at FDP concentrations above 0.1 mM.

FIGS. 7-11 compare cell viability at various concentrations of MSM and FDP. White bars represent vehicle (medium), light grey to dark grey bars represent combinations of MSM and FDP in vehicle, and bars with crossed lines represent data for cells treated with just one substance in vehicle. The best combinatorial effects of MSM and FDP were achieved when FDP was chosen at a concentration of 1 μM and MSM at a concentration of 1 and 0.1 μM.

Example 4—Cell Viability of Primary Chicken Neurons Treated with Rutin

Using the protocol of Example 2, a formulation containing methylsulfonylmethane (0.1 μM), D-fructose 1,6-diphosphate trisodium salt hydrate (1 μM), and rutin (0.001 μM) was tested for cell viability in primary chicken neurons. Vehicle control (medium) and methylsulfonylmethane with fructose 1,6-diphosphate compositions were also tested. The results appear in FIG. 12. The formulation with three active ingredients increased cell viability over vehicle control and MSM+FDP alone, although the increase was not considered statistically significant.

Example 5—Ischemic Challenge

Similar to Example 1, cryopreserved human stem cell derived neurons obtained from PhoenixSongs Biologicals were thawed and plated at a density of 12000 cells per well in a 384-well PDL/laminin coated plate. The neurons were maintained in a humidified environment at 37° C. with 5% CO₂ for six days with periodic media changes before experimental procedures were performed. The media used for the experiment was PhoenixSongs Neural Differentiation Medium and contained methylsulfonylmethane (“MSM”) and fructose 1,6-diphosphate (“FDP”) (specifically, D-fructose 1,6-diphosphate trisodium salt hydrate) with final concentrations of 0.1 μM and 1 μM, respectively, for some cells. The neural differentiation medium optionally with MSM and FDP was spiked with iodoacetic acid (challenge) at 5 μM and was used to challenge the cells for ischemia. Cells with medium and MSM and FDP were dosed in triplicate with 10-point dose response curves to determine if there was any protective effect (improved survival of cells) or toxicity with rutin trihydrate versus the untreated cells.

After 24-hour incubation with rutin trihydrate and iodoacetic acid, the cells were stained with Hoechst and propidium iodide and read on an ArrayScan VTI to determine cell viability. Results were normalized to vehicle (medium alone) and graphed using GraphPad Prism. Statistical significance was determined by performing a one-way analysis of variance followed by a Dunnett test for comparing every test mean to the mean of the media with challenge. The results for rutin trihydrate at various micromolar concentrations appear in FIG. 13. Several concentrations of rutin trihydrate show improved cell viability compared to media with challenge and media/MSM/FDP with challenge, although were not considered statistically significant.

EMBODIMENTS Embodiment 1

A multi-component formulation comprising: methylsulfonylmethane,

fructose 1,6-diphosphate, and

at least one of trehalose, green tea extract, ashwagandha, and rutin.

Embodiment 2

The multi-component formulation of embodiment 1, comprising just one of trehalose, green tea extract, ashwagandha, and rutin.

Embodiment 3

The multi-component formulation of embodiment 2, comprising trehalose.

Embodiment 4

The multi-component formulation of embodiment 2, comprising green tea extract.

Embodiment 5

The multi-component formulation of embodiment 2, comprising ashwagandha.

Embodiment 6

The multi-component formulation of embodiment 2, comprising rutin.

Embodiment 7

The multi-component formulation of any one of the preceding embodiments, wherein the methylsulfonylmethane comprises at least 0.01%, 0.01% to 0.5%, 0.5% to 1.0%, 1.0% to 5.0%, 5.0% to 10%, 10% to 20%, 20% to 50%, 50% to 75%, 75% to 95%, or less than 99% by weight of the formulation.

Embodiment 8

The multi-component formulation of any one of the preceding embodiments, wherein the fructose 1,6-diphosphate comprises at least 0.01%, 0.01% to 0.5%, 0.5% to 1.0%, 1.0% to 5.0%, 5.0% to 10%, 10% to 20%, 20% to 50%, 50% to 75%, 75% to 95%, or less than 99% by weight of the formulation.

Embodiment 9

The multi-component formulation of any one of the preceding embodiments, wherein the trehalose, if present, comprises at least 0.01%, 0.01% to 0.5%, 0.5% to 1.0%, 1.0% to 5.0%, 5.0% to 10%, 10% to 20%, 20% to 50%, 50% to 75%, 75% to 95%, or less than 99% by weight of the formulation.

Embodiment 10

The multi-component formulation of any one of the preceding embodiments, wherein the green tea extract, if present, comprises at least 0.01%, 0.01% to 0.5%, 0.5% to 1.0%, 1.0% to 5.0%, 5.0% to 10%, 10% to 20%, 20% to 50%, 50% to 75%, 75% to 95%, or less than 99% by weight of the formulation.

Embodiment 11

The multi-component formulation of any one of the preceding embodiments, wherein the ashwagandha, if present, comprises at least 0.01%, 0.01% to 0.5%, 0.5% to 1.0%, 1.0% to 5.0%, 5.0% to 10%, 10% to 20%, 20% to 50%, 50% to 75%, 75% to 95%, or less than 99% by weight of the formulation.

Embodiment 12

The multi-component formulation of any one of the preceding embodiments, wherein the rutin, if present, comprises at least 0.01%, 0.01% to 0.5%, 0.5% to 1.0%, 1.0% to 5.0%, 5.0% to 10%, 10% to 20%, 20% to 50%, 50% to 75%, 75% to 95%, or less than 99% by weight of the formulation.

Embodiment 13

A method of making the multi-component formulation of any one of the preceding embodiments, comprising:

combining the methylsulfonylmethane, fructose 1,6-diphosphate, and the at least one of trehalose, green tea extract, ashwagandha, and rutin into a form suitable for administration to a human or animal patient in need thereof.

Embodiment 14

A method of preventing or delaying cognitive decline in a human or animal patient in need thereof, comprising: administering to the patient an effective amount of the multi-component formulation of any one of embodiments 1-12.

Embodiment 15

A method of preventing or delaying Alzheimer's disease or other dementia in a human or animal patient in need thereof, comprising:

administering to the patient an effective amount of the multi-component formulation of any one of embodiments 1-12.

Embodiment 16

A method of treating cognitive decline in a human or animal patient in need thereof, comprising:

administering to the patient an effective amount of the multi-component formulation of any one of embodiments 1-12.

Embodiment 17

A method of treating Alzheimer's disease or other dementia in a human or animal patient in need thereof, comprising: administering to the patient an effective amount of the multi-component formulation of any one of embodiments 1-12.

As previously stated, detailed embodiments of the present invention are disclosed herein; however, it is to be understood that the disclosed embodiments are merely exemplary of the invention that may be embodied in various forms. It will be appreciated that many modifications and other variations stand within the intended scope of this invention as claimed below. Furthermore, the foregoing description of various embodiments does not necessarily imply exclusion. For example, “some” embodiments may include all or part of “other” and “further” embodiments within the scope of this invention. In addition, “a” does not mean “one and only one;” “a” can mean “one and more than one.” 

I claim:
 1. A multi-component formulation to improve neuron viability consisting essentially of: 1.0% to 5.0% by weight methylsulfonylmethane, 20% to 50% by weight fructose 1,6-diphosphate, and one of trehalose, green tea extract, ashwagandha, and rutin, in an amount of at least 0.01% by weight of the formulation.
 2. The multi-component formulation of claim 1, comprising trehalose.
 3. The multi-component formulation of claim 1, comprising green tea extract.
 4. The multi-component formulation of claim 1, comprising ashwagandha.
 5. The multi-component formulation of claim 1, comprising rutin.
 6. A method of making the multi-component formulation of claim 1, comprising: combining the methylsulfonylmethane, fructose 1,6-diphosphate, and the just one of trehalose, green tea extract, ashwagandha, and rutin, into a form suitable for administration to a human or animal patient in need thereof.
 7. A method of treating cognitive decline in a human or animal patient in need thereof, comprising: administering to the patient in need thereof an effective amount of the multi-component formulation of claim
 1. 8. A method of treating Alzheimer's disease or other dementia in a human or animal patient in need thereof, comprising: administering to the patient in need thereof an effective amount of the multi-component formulation of claim
 1. 9. A multi-component formulation to improve neuron viability consisting essentially of: 1.0% to 5.0% by weight methylsulfonylmethane, 10% to 20% by weight fructose 1,6-diphosphate, and one of trehalose, green tea extract, ashwagandha, and rutin, in an amount of at least 0.01% by weight of the formulation.
 10. The multi-component formulation of claim 1, comprising trehalose.
 11. The multi-component formulation of claim 1, comprising green tea extract.
 12. The multi-component formulation of claim 1, comprising ashwagandha.
 13. The multi-component formulation of claim 1, comprising rutin. 